LOCUS       sequence-248261-        5424 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  vector for replacing the promoter of FAS1 coding gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5424)
  AUTHORS   Liu Y, Bai C, Liu Q, Xu Q, Qian Z, Peng Q, Yu J, Xu M, Zhou X, Zhang
            Y, Cai M
  TITLE     Engineered ethanol-driven biosynthetic system for improving 
            production of acetyl-CoA derived drugs in Crabtree-negative yeast.
  JOURNAL   Metab Eng. 2019 Jul;54:275-284. doi: 10.1016/j.ymben.2019.05.001. 
            Epub 2019 May 9.
  PUBMED    31077813
REFERENCE   2  (bases 1 to 5424)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Metab 
            Eng."; date: "2019-07"; volume: "54:275-284. doi"; pages: " 
            10.1016/j.ymben.2019.05.001. Epub 2019 May 9"
COMMENT     Sequence Label: pZ_P-HXT1-FAS1up
FEATURES             Location/Qualifiers
     source          1..5424
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..940
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     CDS             3150..3167
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      3247..3493
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        3508..3921
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3928..3975
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3994..4368
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     terminator      4434..4681
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4450..4468)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(4756..5344)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4836..4855)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag
       61 gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt
      121 tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc
      181 agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta
      241 acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta
      301 tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg
      361 agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct
      421 gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg
      481 ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcggca taccgtttgt
      541 cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct
      601 ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct
      661 ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact
      721 gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat
      781 atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt
      841 actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga
      901 caacttgaga agatcaaaaa acaactaatt attcgaaacg agggtaccca attgattaag
      961 ttcagtgaaa tttcaaaccg ctatacacaa caggacaact ttgagtttag aaaaatccga
     1021 tgtagtgtaa cggctagcac ggtccgcttt caccgggcag acccgggttc gactcccggc
     1081 atcggagttt atttttccat ttcgttcttt agagtattct cctcagcatg cccccctgaa
     1141 tttttccttt tttccatgtg tcccattttt ccactttttt tacagttttc ctcgtgatgt
     1201 taattggcta cacaaaagct gccacacgaa accttaatca cgaaaaacta tacagccttc
     1261 actaatccgt agccccataa tatgttgtcc acgtgctgtt gggtactacc tgtagactct
     1321 cataccccac tccgtctttc tccaacaatt aacgcagtac cgagatttat cagcagactc
     1381 aaattgggca aactctgtat ttttccttgc ccgcataatt tatgggtctc aggcctccac
     1441 gtttcctgtt tacttgaaga atattggctg cggaaaaagt ggtaaggaca accccctttt
     1501 aattggatcc agtttttccg aaatgttccg atccgtacgt catctccgaa gccgtacatt
     1561 ttcactcaat ctacgtagct ttggactcag cgctcctgga attgccagga cagtttactt
     1621 gagttgatat tcccttgtag attgtgtgct tctttttcca aaatttgagg cttcgtttga
     1681 aaagtggaat ctggtcgcta gatcacttca tgcctatttt tcacggaaaa ataagtggta
     1741 ctatgcaccc cttaaaccta aagaaaaacg gaaaaattac cccaaaacct ggtgatgttt
     1801 ttcgcccctt tctttttatc cgagtttttc ttttttcttg tctgccaaat tcctctcctg
     1861 accttagcgt ccccggaaaa aattaactac ttaaggaccg aatgagcccc agcttttccc
     1921 cttctcttca ttattcccca taatataata tgagtgctac atccggagtt gtcaacaggc
     1981 cattggtgtt aaaccatggc tcaatcgagt caaccatctt aatcccaacc acagaatatc
     2041 atttctacca aacgttactg gaagggttca ggaagtcatt gccacaagtt accgagggat
     2101 ttgctgacga cgacgagccg agctccaagg cagaacttct gatgaagttt ttgggttaca
     2161 ttgtccaatc tggagtctcc aatcagcaag agcaattagc tgctgcaaag ttggttttga
     2221 atgagttcga gtcccgtttc cttcaaggat tgaacttgca ctcttatgct gccattttgc
     2281 ttaagagcga gacctttccc acaacattat tgaaaatcaa agaaaactta atcaagaact
     2341 actacctggg aagagccctt gtctacctgc caggtcagcg tggccttgta taccctccct
     2401 cggcattact taatgctgga aagtccggct ccgctcagat ctacgccata tttggaggtc
     2461 aaggtaacac cgatgactat ttcgaagaac ttagagatat ttaccacatc taccaaggtc
     2521 tcgtgtcaga cttcgttacc aaggcccagt tgaagttaca agagctcatc agaaccaccc
     2581 ctgaaactga cagaatatat acacagggtc tagacctgat caattggctg gaaaacaagg
     2641 acaagactcc tgaccaacaa caattgctat ccattccaat gtcatgtcct ctaatttgtg
     2701 ttatccaact gtgtcattac attgtcactt gccgcatctt aggtattaca cctggtcaac
     2761 tcagagattc cttgaaaggt actacaggtc actcccaagg tctggtcacc gctgtggtcg
     2821 tttcttctgc tgattcctgg gaatcctttg agaaattggc gcttcaggca gttgaattca
     2881 tgttctacat tggtgttaga ggactacaaa cctatccaaa cacctcttta cctccatcca
     2941 tcgtccaaga ttccgaagaa aatgccgagg gaactccttc tcccatgctt tcagttagag
     3001 acctttccta tgaccaattg gtcaagtttg tcaacgaaac gaaccagcac ttgcctgagg
     3061 ccaaacatat cgacatctct ttaataaatg ggcccagaaa tgtcgtcttg acaggccctc
     3121 ctcaatcact ttacggtttg aatctgaacc atcatcatca tcatcattga gtttgtagcc
     3181 ttagacatga ctgttcctca gttcaagttg ggcacttacg agaagaccgg tcttgctaga
     3241 ttctaatcaa gaggatgtca gaatgccatt tgcctgagag atgcaggctt catttttgat
     3301 acttttttat ttgtaaccta tatagtatag gatttttttt gtcattttgt ttcttctcgt
     3361 acgagcttgc tcctgatcag cctatctcgc agctgatgaa tatcttgtgg taggggtttg
     3421 ggaaaatcat tcgagtttga tgtttttctt ggtatttccc actcctcttc agagtacaga
     3481 agattaagtg agaccttcgt ttgtgcggat cccccacaca ccatagcttc aaaatgtttc
     3541 tactcctttt ttactcttcc agattttctc ggactccgcg catcgccgta ccacttcaaa
     3601 acacccaagc acagcatact aaatttcccc tctttcttcc tctagggtgt cgttaattac
     3661 ccgtactaaa ggtttggaaa agaaaaaaga gaccgcctcg tttctttttc ttcgtcgaaa
     3721 aaggcaataa aaatttttat cacgtttctt tttcttgaaa attttttttt ttgatttttt
     3781 tctctttcga tgacctccca ttgatattta agttaataaa cggtcttcaa tttctcaagt
     3841 ttcagtttca tttttcttgt tctattacaa ctttttttac ttcttgctca ttagaaagaa
     3901 agcatagcaa tctaatctaa gggcggtgtt gacaattaat catcggcata gtatatcggc
     3961 atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag tgccgttccg
     4021 gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg gctcgggttc
     4081 tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc
     4141 atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc
     4201 ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc
     4261 tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc
     4321 gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca cgtccgacgc
     4381 ggcccgacgg gtccgaggcc tcggagatcc gtcccccttt tcctttgtcg atatcatgta
     4441 attagttatg tcacgcttac attcacgccc tccccccaca tccgctctaa ccgaaaagga
     4501 aggagttaga caacctgaag tctaggtccc tatttatttt tttatagtta tgttagtatt
     4561 aagaacgtta tttatatttc aaatttttct tttttttctg tacagacgcg tgtacgcatg
     4621 taacattata ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg ctttaatttg
     4681 caagctggag accaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4741 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4801 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4861 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4921 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4981 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5041 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5101 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5161 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5221 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5281 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5341 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5401 ttaagggatt ttggtcatga gatc
//
